Studies On Preparation And Quality Control Of Gegenqinlian Injection

Gegenqinlian was a classic preparation of Shanghan Lun which widely used to cure damp-heat syndrome. At present, Gegenqinlian had been prepared in various kinds of dosage forms such as tablets, Oral, particle, drop pill, micro-pills, patch, and colon-targeted capsules. However, no preparation was applied on animal except decoction.In view of active-ingredients of Gegenqinlian were so clear that the orthogonal design can be used to evaluate the method of extraction, purification, preparation and quality control.1. The Optimum extract condition was:Radix Pueraria, Radix Scutellaria and Radix Glycyrrhizae were extracted together in terms of 11 times water of medical material, boiled 2 times and each time 1.5 hours. Rhizoma Coptis was extracted alone in the same way. The alcohol was selected as the Optimum purification method.2. The Optimal method of preparation was:4%HP-β-CD,1.2%tween-80 and 30% propylene glycol were added in the mixed decoction, which would be prepared to Gegenqinlian Injection after Sterilization.3. The standard quality was established on studies of Gegenqinlian Injection. It was identified by TLC from which we can see that the spots were clear and that the recurrence was as expected. Puerarin was detected by Silca gel-H, the mobile phase consisted of chloroform-methanol-water (5:1.5:0.25) and detection was performed at 254nm. Baicalin was examined by Silica gel-H, the mobile phase was ethyl acetate-acetone-n-butanol-acetic acid-water (6:3:1:1:3) and detected by 1% chloride ferric solution that dissolved by alcohol. Berberine hydrochloride was examined in the mobile phase that consisted of n-butanol-acetic acid-water (6:1:2), and the detection wavelength was 366nm.A reversed-Phase HPLC method was established to determine the content of puerarin, baicalin and berberine in Gegenqinlian injection.The result showed the calibration curve of Puerarin was linear in the range of 0.7-3.2μg/ml.The regression equation was y=47303x+2.4455×104 and the correlation coefficient was 0.9999(n=5).The average recoveries is 96.0%(RSD=1.1). The calibration curve of Baiealin was linear in the range of 4.3-12.9μg/ml.The regression equation was y=33471x+2.5156x105 and the correlation coefficient was 0.9999(n=5).The average recoveries is 95.6%(RSD=1.2). The results showed the calibration curve of Berberine was linear in the range of 4.0-20.0μg/ml.The regression equation was y =12012x+2.6244x105 and the correlation coefficient was 0.9999(n=5).The average recoveries is 96.4%(RSD=0.9). The content of puerarin, baicalin and berberine were controlled to 2.086mg/ml,2.573mg/ml and 6.836mg/ml as qualified.4.In vitro bacteriostatic test showed the MIC of Gegenqinlian Injection against S.aureus was 14.125μg/ml, while S.enterica and E.coli were 56.5μg/ml,226μg/ml。The content of Puerarin in Gegenqinlian Injection was controlled to 2.086mg/ml as qualified. The Baiealin was 2.573mg/ml and the Berberine was 6.836 mg/ml.